Supplementation of biotin to sperm preparation medium enhances fertilizing ability of spermatozoa and improves preimplantation embryo development

Abstract

Purpose Motility of spermatozoa helps not only in planning the type of infertility treatment but also directly reflects the success rate in assisted reproductive technology (ART). Previously, biotin, a water-soluble vitamin, has been shown to increase the motility and longevity of cryopreserved human spermatozoa. The present study was designed to understand the molecular basis of the beneficial effects of presence of biotin in sperm wash medium on early embryo development. Methods The effect biotin supplementation to sperm wash medium on the sperm parameters were assessed in swim-up fraction of normozoospermic and asthenozoospermic ejaculates collected from infertile men. Fertilization and early embryo development was studied using Swiss albino mice. Results Even though both biotin and pentoxifylline (PTX) enhanced the motility of spermatozoa from normozoospermic and asthenozoospermic samples, biotin group exhibited higher in vitro survival. Using mouse model, we observed that presence of biotin or PTX in sperm wash medium improved the fertilization rate and blastocyst rate compared to control. Blastocysts from these groups had significantly higher total cell number (P < 0.01) and lower apoptotic index. In silico target prediction revealed that GTPase HRas (HRas), tyrosine-protein phosphatase nonreceptor type 1 (PTP1B), and glucokinase are the probable targets for biotin. Solution-state Nuclear Magnetic Resonance (NMR) studies confirmed that biotin interacts both with human HRas and PTP1B. Conclusion Our results indicate that presence of biotin in sperm wash medium can improve the fertilization potential and preimplantation embryo development and can be considered as a safe alternate to PTX.

Current Insights and Latest Updates in Sperm Motility and Associated Applications in Assisted Reproduction

“…At physiological concentrations, [178] Fallopian tubal cells [225] Vero cells [226] Hormone and growth factors Insulin and leptin [177] Platelet activating factor [227,176] Follicular fluid [228,229] Progesterone [230] Leukemia inhibiting factor [231] Thyroxine [173] Relaxin [175] Müllerian inhibiting substance [232] Human chorionic gonadotropin [174] Bradykinin [233] High-energy molecules and prostaglandins Creatine phosphate [234] cAMP, cyclic adenosine monophosphate; 8-MeIBMX, 8-methoxymethyl-3-isobutyl-1-methylxanthine; IBMX, 3isobutyl-1-methylxanthine; PDE, phosphodiesterase bradykinin, angiotensin I, II, and III, and acetylcarnitine exhibited a direct stimulating effect on sperm motility in vitro [180]. Various vitamins and antioxidants were shown to improve sperm motility and longevity by reducing in vitro oxidative stress [181]. Few herbal medicines, rich in antioxidants such as Tribulus terrestris extract [182,183] and Mondia whitei [184], have been reported to enhance sperm motility in vitro.…”

Early prepubertal cyclophosphamide exposure in mice results in long-term loss of ovarian reserve, and impaired embryonic development and blastocyst quality

“…In vitro fertilization and embryo culture were carried as described earlier [20]. OCCs from the oviduct were released into pre-warmed Earl's balanced salt solution (EBSS, E2888, Sigma Aldrich, USA) supplemented with 0.1% bovine serum albumin (BSA).…”

Does in vitro application of pentoxifylline have beneficial effects in assisted male reproduction?

“…Pentoxifylline belongs to the family of phosphodiesterase inhibitors (PDE) that leads to an increase of the cyclic adenosine monophosphate (c-AMP) levels. This leads to an increased number of acrosome reactions as well as sperm movement characteristics (Esteves, Spaine, & Cedenho, 2007;Nassar et al, 1998;Salian et al, 2019). PTX also shows antioxidant characteristics (Chehab, Madala, & Trussell, 2015) and, via prohibiting xanthine oxidase, leads to a reduction of the levels of intracellular reactive oxygen species (ROS) produced by defective spermatozoa, also diminishing lipid peroxidation (McKinney, Lewis, & Thompson, 1996).…”

“…Furthermore, enhanced lipid peroxidation reduces membrane fluidity, leading to diminished sperm motility and deficiency of sperm essential functions, such as the acrosome reaction (Alahmar, 2019). Both the antioxidative activity and radical scavenging characteristics of PTX have been shown to improve the likelihood of acrosome reactions (Esteves et al, 2007;Salian et al, 2019;Williams & Ford, 2005) and hyperactivation of spermatozoa (Kay, Coutts, & Robertson, 1993;Nassar et al, 1998;Pang, Chan, & Lu, 1993). Treatment of sperm samples with PTX is done in two different ways: 1, with addition of certain volume (depending on the final concentration) of PTX stock to the resuspended sperm pellet (Amer, Metawae, Hosny, & Raef, 2013;Salian et al, 2019); 2, addition of PTX solution that has been prepared at final concentration to the microdroplets of an ICSI Petri dish (Navas et al, 2017) (Figure 2).…”